Fluorescence Digital Image Gallery

Embryonic Swiss Mouse Fibroblast Cells (3T3)

The progress made in the cultivation of human epidermal tissues enabled by the 3T3 cell line was vital in the development of improved treatments for burn victims. However, before cultured keratinocytes could be utilized for therapeutic purposes, it was necessary to find a method of detaching the epithelial tissue in sheets while eliminating most of the 3T3 cells utilized for culturing. Once this was achieved, cultivated keratinocytes began being utilized experimentally to treat mice, and then humans, who had suffered loss of epithelia with considerably successful results. The cells were found to be capable of regenerating epidermal tissue in a relatively short period of time, although the production of the anchoring networks needed to connect this tissue with underlying tissue layers was significantly less rapid. Nevertheless, keratinocytes grown with the help of 3T3 cells are now regularly utilized in burn centers around the world for skin grafts.

The culture of 3T3 cells featured in the digital image above was immunofluorescently labeled with primary anti-tubulin mouse monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to the cyanine dye, Cy3. In addition, the cells were simultaneously probed for DNA with the ultraviolet-absorbing probe DAPI, and for the cytoskeletal filamentous actin network with Alexa Fluor 488 conjugated to phalloidin. Images were recorded in grayscale with a QImaging Retiga Fast-EXi camera system coupled to an Olympus BX-51 microscope equipped with bandpass emission fluorescence filter optical blocks provided by Omega Optical. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles.